Preparation of enteric coated digestive enzyme compositions

ABSTRACT

Improved enteric coated digestive enzyme-containing compositions which are capable of withstanding hours of exposure to gastric fluids while protecting the biological activity of the enzymes and thereafter releasing the digestive enzymes in their biologically active state within 5 to 30 minutes after being exposed to intestinal fluids, these compositions comprising (a) an enzyme concentrate in (b) a binder system comprising at least about 0.5 wt. %, preferably about 1 to about 10 wt. % (based on the weight of the binder system plus enzymes) of (i) a binder, preferably selected from the group consisting of polyvinylpyrrolidone, microcrystalline cellulose (Avicel), cellulose acetate phthalate, methylcellulose and alginic acid, and preferably (ii) from about 0.1 to about 10 wt. % of a stabilizer, preferably selected from the group consisting of calcium carbonate, polyvinylpyrrolidone, cellulose acetate phthalate, methylcellulose, alginic acid, starch and modified starches, e.g., carboxymethyl starch (Primojel); and (c) from about 0.1% to about 30 wt. %, based on the weight of the total composite (enzyme plus binder system plus disintegrant) of a disintegrant, preferably selected from the group consisting of citric acid, sodium carbonate, sodium bicarbonate, calcium carbonate and other suitable carbonates, alginic acid, starch and modified starches, e.g., carboxymethyl starch (Primojel) are prepared by a process in which the presence of water is avoided and which includes the step of blending enzyme, binder and disintegrant in the presence of a selected inert solvent as well as the subsequent coating of the resulting enzyme/binder/disintegrant composite with from about 2.5% to about 10% by weight, based on the weight of the enzyme/binder/disintegrant composite, of a gastric juice insoluble, intestinal juice soluble, non-porous, pharmaceutically acceptable enteric coating polymer.

CROSS REFERENCE TO RELATED APPLICATIONS

This is a continuation-in-part of my copending application, Ser. No.585,621, filed June 10, 1975, now abandoned.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to enzyme compositions for ingestion by a mammalhaving a digestive disorder which is caused by an enzyme deficiency orwhich, in any event, can be alleviated by enzyme supplements. Moreparticularly, the invention relates to improved enteric coatedenzyme-containing compositions for ingestion by a mammal, as well as tomethods for making such a composition.

2. Description of the Prior Art

It is well documented in the literature that exogenously administeredpancreatic enzymes from animal sources can remedy the enzyme deficiencycaused by various diseased states of the pancreas, e.g., pancreatitis,pancreatectomy, cystic fibrosis, etc. Fewer data exist for enzymes fromplant and microbial sources.

Pancreatic enzymes are active under near neutral and slightly alkalineconditions. Under gastric conditions, i.e., in the presence of acid andpepsin, most of the enzymes are irreversibly inactivated with resultingloss of biological activity. Therefore, it is imperative that theexogenously administered enzymes be protected against gastricinactivation and remain intact during their transit through the stomachinto the duodenum.

While the transit of the enzymes intact through the stomach isessential, it is another requirement for maximum efficacy that theenzymes be released in the duodenum within 5 to 30 minutes, sincedigestion by pancreatic enzymes and absorption of the metabolites takeplace primarily in the upper segment of the intestine, i.e., duodenumand upper part of the jejunum.

The normal pancreas, in reponse to food stimulation, gradually releasesthe digestive exzymes, mostly in their inactive precursor form, into theduodenum. Some of the released zymogens (inactive precursors) are firstactivated by enterokinase to form active enzymes, e.g., trypsin fromtrypsinogen. The newly activated trypsin in turn generates more activeenzymes in the duodenum by an autocatalytic mechanism. Simultaneously,the activated enzymes are thoroughly mixed with the arriving food fromthe stomach, and digestion ensues. This process takes place as long asfood is pumped from the stomach into the duodenum.

Pancreatic enzymes have been used for the past seventy years to treatvarious digestive disorders. The early clinical results were variable.With time it became apparent that some of the poor clinical responseswere due to gastric inactivation of the exogenously administeredenzymes. A revived interest in enzyme-containing digestive aids occurredin the late 1950's and early 1960's, with the development of acid stableenteric coatings. Thus, it was believed that the detrimental effects ofgastric acidity on the enzymes could be avoided by the use of suchenteric coatings, and more effective enzyme therapy thus made possible.A great variety of enteric coated enzyme-containing digestive aids weremarketed in this period. However, most of these products contained lowlevels of active enzymes, often too low to effectively treat many enzymedeficiency-related conditions. Many of these products were particularlydeficient in lipase.

Moreover, the coatings generally failed to protect the enzymes againstgastric inactivation or to release them in an activatable state in theduodenum. Thus, most were permeable to gastric acid and many failed todisintegrate in the duodenum under neutral conditions within areasonable time, from the point of view of being available in properconcentration in active form at the time when food which has passedthrough the stomach is present in the duodenum and upper jejunum.

Because of these known defects in the coatings as well as the low levelsof enzyme activities of prior art digestive enzyme-containingcompositions, it has long remained a desired goal to develop a highlyactive enzyme-containing digestive aid composition that would preventgastric acid and pepsin inactivation of the enzymes upon passage throughthe stomach, and, after transit from the stomach into the duodenum,would release the enzymes in a reproducible, i.e., predictable, mannerwithin minutes in their biologically active state.

In a pancreatic deficient state the normal physiological conditions ofdigestion (i.e., gradual release of pancreatic zymogens into theduodenum, their activation by enterokinase and their even mixing withthe incoming food from the stomach) in the duodenum are absent orgreatly impaired. Exogenous administration of the enzymes in largetablets can aggravate the already existing abnormal physiological statebecause large amounts of enzymes may be released into small,concentrated areas. This in turn could result in irritation and damageto the intestinal lining. Furthermore, any asynchrony that may existbetween the arrival of food and of the tablets from the stomach into theduodenum will further reduce the possibility that normal digestion cantake place under this condition. A tablet can arrive in the duodenum tooearly or too late with respect to arrival of the food. This results inunpredictable response and poor digestion.

As can be seen from the foregoing, the desiderata for a successful,highly acceptable digestive enzyme composition, which have notheretofore been satisfactorily provided by any one product, include: (1)providing a properly enteric coated composition which enables deliveryof the exogenously administered enzymes intact through the stomach intothe duodenum; (2) ensuring the release of the protected enzymes, in abiologically active state, into the duodenum within minutes after thepassage of the compositions from the stomach into the duodenum byproviding an enteric coating which will promptly dissolve in intestinaljuices, and controlling the chemical composition, physical form and sizeof the digestive enzyme composition to promote rapid disintegration upondissolution of the enteric coating; and (3) recreating the physiologicalconditions for digestion that exist in the duodenum with a normalfunctioning pancreas, by (a) providing the composition in small unitsize form to promote even mixing thereof with the food in the stomach sothat the enzymes are gradually released into the duodenum and areuniformly dispersed throughout the food arriving from the stomach, and(b) assuring that sufficient activators such as co-lipase are present toprovide for the patient maximum benefit from his own naturally producedenzymes as well as those that are administered exogenously.

SUMMARY OF THE INVENTION

The present invention provides compositions, as well as methods fortheir preparation and use, that promote the delivery in a reproduciblemanner into the upper intestine, in a bio-available form, of exogenouslyadministered digestive enzymes, as well as their release in acontrolled, biologically acceptable manner. By virtue of this invention,a highly effective system is provided that greatly improves thenutritional and psychological well-being of enzyme deficient patients.With the regular use of this system, food digestion is aided. Inaddition, maldigestion due to enzyme deficiency as well as bacterialfermentation of undigested food in the lower intestine are prevented.

The compositions of the present invention comprise an enzyme concentratein a binder selected from the group consisting of polyvinylpyrrolidone,microcrystalline cellulose, cellulose acetate phthalate, methylcelluloseand alginic acid; and from zero to about 10 wt. % of a stabilizerselected from the group consisting of calcium carbonate,polyvinylpyrrolidone, cellulose acetate phthalate, methylcellulose,starch and modified starches and alginic acid; and from about 0.1% toabout 30 wt. %, based on the total weight of the composite, of adisintegrant selected from the group consisting of citric acid, sodiumcarbonate, sodium bicarbonate, calcium carbonate, starch and modifiedstarches, and alginic acid; the foregoing enzyme/binder/disintegrantcomposite being coated with from about 2.5% to about 10% by weight,based on the weight of the enzyme/binder/disintegrant composite, of anon-porous, pharmaceutically acceptable enteric coating polymer, wherebyto provide an enteric coating which is insoluble in the pH range of fromabout 1.5 to about 5 normally existing in mammalian gastric fluids, butsoluble at a pH of from about 6 to about 9, the normal pH range formammalian intestinal fluids, so that said finished composition iscapable of withstanding at least about one hour, preferably two hours,of exposure to mammalian gastric conditions, but will dissolve withinabout 5 to about 30 minutes in intestinal juices of mammals.

In one preferred embodiment, the present invention provides compositionswhich comprise a pancreatic enzyme mixture, each milligram of whichcontains at least about 75 N.F.* units of protease, at least about 75N.F. units of amylase, at least about 10 N.F. units of lipase,preferably at least about 5 International Units (IU) of ribonuclease,and an effective amount of co-lipase in an enteric coated composition asdescribed in the preceding paragraph.

In accordance with a preferred feature of the present invention, theenteric coated enzyme compositions are provided in the form of "smallbeads", spherical particles having diameters in the preferred range offrom about 8 to about 14 mesh. In theory, the smallest beads give thebest distribution of enzymes in the food. However, extremely small beadsare more difficult to form since they require an extremely finelypowdered enzyme to form true spheres, whereas excessive milling of theenzyme powder irreversibly inactivates some of the enzymes. Extremelysmall enzyme beads also require a larger percentage of enteric coatingdue to the greater surface area. Therefore, although smaller than 14mesh and larger than 8 mesh beads are suitable, the most practical sizerange is 8 to 14 mesh. Particularly preferred are beads in the 10 to 12mesh range. As used herein, the expression 8 mesh means that the beadwill be caught on a screen having a U.S. Series designation of 8 (whichhas square sieve openings of 2.38 mm), but pass through a screen havinglarger openings. Correspondingly, 14 mesh beads are caught on a screenhaving a U.S. series designation 14 (or openings of 1.68 mm), but passthrough a screen having larger openings. For convenience ofadministration to the patient, these beads may be provided in gastricjuice soluble capsules. Each capsule may suitably contain from about 250to about 600 mg. of beads, preferably from about 425 to about 475 mg.

As an alternative, the more commonly used tablet form may be employed.In this event, however, it is preferred to limit the size of the coatedtablet to the smallest feasible size in order to enhance the uniformityof distribution of the compositions of the present invention within thefood passing from the stomach into the duodenum. Another advantage ofemploying a large number of smaller tablets is that inactivation of theenzymes in one tablet in the stomach due to an inadvertent flaw in theenteric coating will result in the loss of only a relatively smallpercentage of the total enzyme administered. For convenience, thesetablets are preferably also administered in gastric juice solublecapsules, each capsule typically containing a plurality of smalltablets.

In general, advantages of the bead form over the tablet form includeprovision of more uniform distribution of enteric coated enzymecomposition throughout the food in the stomach due to the smaller sizeof the beads, the ability to readily obtain more uniform entericcoatings due to the spherical shape of the beads, and greater ease ofdisintegration in the intestines, since the poorer disintegrationcharacteristics and loss of enzymatic activity caused by the pressuresof compaction required to obtain a coherent tablet may be avoided inproducing the spheres.

In accordance with another aspect of this invention, the compositions ofthe invention are prepared by a process which comprises granulating theenzyme or enzyme mixture, together with the binder and disintegrant, aswell as the stabilizer, if present as a separate entity, inapproximately 600-700 ml./kg. (of solids) of an inert solvent selectedfrom the group consisting of isopropanol, methylene chloride, dioxane,tetrahydrofuran and acetone.

In a particularly preferred process in accordance with the presentinvention, the enzyme or enzyme mixture and disintegrant are mixed inthe dry state, the binder (together with the stabilizer if any) isdissolved in a solvent as defined below to form a binder solution, andthe composition is formed into beads having a diameter in the range offrom about 8 to about 14 mesh by dusting said dry blend over nonpareilseeds tumbling and flowing in a coating pan, said seeds having beenwetted with said binder solution, with periodic addition of bindersolution to maintain the particles in a wetted but free-flowing state,until the seeds have been built to uniform spherical particles havingdiameters predominantly in said range of from about 8 to about 14 mesh.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

The compositions of this invention include a digestive enzymeconcentrate. In a presently preferred embodiment, they include theenzymes amylase, protease and lipase, together with co-lipase, andpreferably also include ribonuclease. They also include at least onebinder selected from the group consisting of cellulose acetatephthalate, polyvinylpyrrolidone, microcrystalline cellulose, alginicacid and methylcellulose and at least one disintegrant selected from thegroup consisting of sodium carbonate, sodium bicarbonate, citric acid,starches and modified starches (e.g., Primojel, Sta-Rx),microcrystalline cellulose, and alginic acid. As is known, when acarbonate is used as a disintegrant, it is used in combination with amild acid, such as citric acid or tartaric acid. Preferably, they alsoinclude a separate stabilizer selected from the group consisting ofcalcium carbonate, polyvinylpyrrolidone, cellulose acetate phthalate,methylcellulose, starch and modified starches. It will be noted that thesame material, e.g., polyvinylpyrrolidone, can act both as the binderand the stabilizer.

The principal active agents in the compositions of this invention, theenzymes, include such pancreatic enzymes as (I) the proteases, forexample, Trypsin, E.C. (Enzyme Commission Number) 3.4.4.4; Chymotrypsin,E.C. 3,4,4,5; Chymotrypsin B, E.C. 3,4,4,6; Pancreatopeptidase E, E.C.3.4.4.7; Carboxypeptidase A, E.C. 3.4.2.1; and Carboxypeptidase B, E.C.3.4.2.2; (II) the lipases, for example, Glycerol ester hydrolase(Lipase), E.C. 3.1.1.3; Phospholipase A₂, E.C. 3.1.1.4; and Sterol esterhydrolase, E.C. 3.1.1.13; (III) the nucleases, for example,Ribonuclease, E.C. 2.7.7.16 and Deoxyribonuclease, E.C. 3.1.4.5; and theamylase, α-Amylase, E.C. 3.2.1.1. Among suitable digestive enzymesderived from plant sources are papain, E.C. 3.4.4.10; Chymopapain, E.C.3.4.4.11; Bromelain, E.C. 3.4.4.c; Ficin, E.C. 3.4.4.12; and β-Amylase,E.C. 3.2.1.2. Suitable digestive enzymes derived from microbial sourcesinclude β-Galactosidase (Lactase), E.C. 3.2.1.23; Cellulase, E.C.3.2.1.4; Subtilopeptidase A., E.C. 3.4.4.16; and Aspergillopeptidase A,E.C. 3.4.4.17.

Generally, the enzymes are available in powder or crystalline form,typically as concentrates of pancreatic enzymes (protease, amylase,lipase and, preferably, ribonuclease) derived from animal sources (hog,sheep and bovine). However, plant and microbial derived enzyme systemscan also be used, if desired. Desirably, co-lipase is also included. Toassure proper therapeutic effectiveness for pancreatic enzyme deficientpatients, the initial enzyme activities per milligram should be at leastthose set forth above. In any event, for properly controlled therapy, itis important both to know the initial enzyme activities and to be ableto predict the corresponding activity upon release in the intestinaltract. It should be understood that, as used herein, the term "enzyme"includes not only the already activated form but also the zymogenprecursor which is capable of being transformed into the active form inmammalian intestinal fluid.

Suitable binders for granulation include polyvinylpyrrolidone (PVP)(Plasdone), microcrystalline cellulose (Avicel), cellulose acetatephthalate, and methylcellulose (Methocel). The binders are generallypresent in amounts of about 0.5% to about 10% by weight, preferably fromabout 1% to about 5% by weight. While greater percentages of bindercould be used, I have found no advantage in exceeding 10% by weight.

Suitable stabilizers and disintegrants have been listed above andinclude, generally, those known to the art. The preferred weight ratioof disintegrants to enzymes is between about 2 and 15%, although betweenabout 0.5% and 30% by weight may suitably be used.

The compositions of the invention may be prepared in tablet or bead formin accordance with the techniques described below. While each of theseprocesses is analogous to processes employed heretofore in thepreparation of other compositions, each is unique as applied to enzymepreparations. In particular, the discoveries that certain inert solventsmay be used with the enzymes, as discussed below, as well as theparticular parameters herein described in connection with the use of thesolvents, are important features of the present invention.

(a) Granulation

In accordance with one aspect of the present invention, and as apreliminary step for both the production of tablets and the productionof beads by segmentation or marumerization, the enzyme powder, thebinder system and the disintegrant are blended together and granulatedin an inert solvent as described in Remington's Practice of Pharmacy(RPP), XII pp. 445-449 (1961), and then extruded into segments throughopenings in the range of from about 0.5 to about 2 mm in diameter,preferably from about 0.8 to about 1.2 mm. These segments have a solventcontent of about 47 to 55% by weight. Then the segments are furtherprocessed to their final form (e.g., tablet or bead) as described below.

In an alternative procedure of granulation, which has been employed forexample with the enzyme blend of Example 6, the binder, e.g.,polyvinylpyrrolidone, and the disintegrant, e.g., citric acid, are firstdissolved in the inert solvent, e.g., isopropanol, and then slowly addedto the enzyme mixture. Similarly to the previously described procedure,and as discussed below, the proper amount of solvent used for thegranulation was found to be between about 600 and 700 ml/kg of enzymeblend. The advantage of this modified procedure over the previous one isthat a more even distribution of the binder and the disintegrant areobtained. This in turn results in a more uniform product with fasterdisintegration and longer shelf-life (stability).

(b) Tablet Preparation

Tableting of the extruded and screened segments to form small tablets ofconvenient size, e.g., 4.8 mm. diameter and 4 mm. thickness, can beachieved by conventional tableting procedures at relatively lowcompression pressures and at temperatures of about 15° to 30° C. It hasbeen observed that high compression pressures, which produce hardtablets having slow disintegration rates in intestinal juices, alsoresult in elevated compression temperatures (i.e., 35° C and above).This causes a loss in biological activity. Preferably, the friability ofthe tablets should result in less than a 1% loss in 4 minutes understandard testing procedures. The hardness should be about 2 to 5kilograms, preferably about 2.5 to 3.5.

In accordance with one aspect of the invention, the (uncoated) tabletsare of relatively small size, having a diameter between about 2 and 7mm., and a thickness between about 2 and 5 mm.

(c) Bead or Small Sphere Preparation

In accordance with another aspect of the invention, the segments areformed into beads having an average size in the range of from about 8 toabout 14 mesh, preferably from about 10 to about 12 mesh.

One method of spheronization, or creation of beads, is to process thesegments directly in a Marumerizer (see "A New Technique for theProduction of Spherical Particles" by A. D. Reynolds, ManufacturingChemist & Aerosol News, June 1970, p 40) for a period of from about 30to about 75 seconds at a temperature of about 20° C. This procedure,while satisfactory, results in a wide range of particle sizes with arelatively low yield of the desired beads. In general, longermarumerization times result in more compact particles, which, by virtueof their greater compactness, require longer times for disintegration.

(d) Spherical Beads by a Non-Pareil Process

In accordance with yet another aspect of the invention, there isprovided an alternative, preferred, procedure for the formation of smallspheres or beads. This is based on the discovery that, under carefullycontrolled conditions as described below, larger spherical particles ofdigestive enzyme compositions can be obtained from small sphericalparticles (nonpareil seeds, typically sugar, having a mesh sizegenerally in the range of from about 20 to about 32) by slowly dustingthe enzyme/disintegrant composite over the tumbling and flowingnonpareil seeds wetted with the solvent containing the binder dissolvedtherein (binder solution) in a conventional coating pan, until the seedsbuild up in size to spherical particles having diameters in the range offrom about 8 to about 14 mesh. In carrying out this process, the enzymes(see e.g., Example 9, Phase 1) are preblended with a disintegrant,preferably carboxymethyl starch (Primojel), in a Hobart mixer for 10 to15 minutes. In a separate container, the binder/stabilizer, e.g.,polyvinylpyrrolidone (PVP), blend is dissolved in 725 ml of a solvent,preferably isopropanol per kilogram of blend. The nonpareil sugar seedsare prewetted with the isopropyl alcohol-PVP solution (the degree ofwetness being sufficient to promote adhesion of enzyme disintegrantmixture to the wetted sugar particles but not so great as to promoteagglomeration of the beads), after which the preblended mixture ofpancreatic enzymes and disintegrant is slowly sprinkled over thetumbling nonpareil sugar seeds in a conventional coating pan (e.g., asavailable from the Manesty Co.). The sequence of the above steps isrepeated until the seeds are built up into beads of the desired sizes.

The above procedure results in high yields of uniform particle size,approximately 90% of the particles being within the range of 8 to 12mesh, with the preservation of 90 to 95% of the biological activity. Bycomparison, during a conventional tableting operation under theconditions of the present invention, only about 70 to 90% of thebiological activity is generally preserved.

e. Solvent

The solvent serves as the wetting agent. The term "inert solvent" asused herein means that the solvent does not have a deleterious effect onthe biological activity of the enzymes nor adversely react with anyother component of the composition of this invention.

Suitable inert solvents for use as wetting agents in theblending/granulating step include isopropanol, methylene chloride,dioxane, tetrahydrofuran, and acetone. These may be used singly or inadmixture with each other. (Methanol and ethanol, at least when used asthe only solvents, have been found to be detrimental to the biologicalactivity of the preparation under the foregoing conditions used forgranulation, i.e., these solvents tend to destroy the biologicalactivity of the enzymes.) The preferred solvents for granulation werefound to be isopropanol, methylene chloride and dioxane. The mostpreferred solvent is isopropanol. (See Table III.)

Proper wetting of the enzyme powder with the solvent, e.g., isopropanol,for the granulation and for the subsequent manufacturing operation isimportant to achieve, and is dependent on the solvent concentration.Overwetting results in poor segmentation in the extruder and stickinessin the marumerization steps. Overwetting produced similar results whenthe nonpareil seeds were used, i.e., the beads stuck to the coating panand they formed multiplets or agglomerates.

The proper amount of isopropanol for granulation was found to be betweenabout 600 and about 700 ml/kg of enzyme blend. Best results wereobtained with 625 to 675 ml of isopropanol per kg of enzyme blend. Inthe nonpareil process, evaporation is relatively rapid due to highsurface area, so that proper wetting is a matter of continuedsurveillance, with periodic addition of binder solution as needed.

I have also found that water must be meticulously avoided during anyphase of the processing operation since the presence of water even insmall amounts is detrimental to the biological activity of the enzymes.Therefore, it is also important that incidental moisture contentresulting from normal processing of raw materials as well as thehumidity of the air in the facility where the manufacturing of theenzyme composition is carried out to be kept as low as possible.

f. Enteric Coating

I have discovered that in order to obtain an enteric coating polymerhaving the necessary properties to survive gastric conditions for atleast about 1 hour, preferably 2 hours, and readily disintegrate in theduodenum under neutral to alkaline pH, the enteric coating polymer mustcomprise at least about 3 to 3.5% by weight of the uncoated beads,whereas about 2.5 wt. % of enteric coating polymer (based on the weightof the uncoated tablets) is sufficient for the tablets. To assureadequate protection against gastric conditions, particularly in the caseof tablets on which it is more difficult to obtain a uniform coating,the thickness of coating at the edges generally differing from that atthe central portion, I prefer to use amounts of about 3 to about 8% w/wof enteric coating polymer. While greater amounts of enteric coatingpolymer could be used, there would be no advantage to exceeding about10% by weight, as the thicker the coating, the slower the dissolution inthe normal intestinal environment. The most preferred range of entericcoating polymer is between about 4.5 and 8% w/w for the beads; andbetween about 3.5 and about 5% for the tablets.

In one preferred method of carrying out the coating operation, separatesolutions of 1%, 2% and 4% concentration are prepared by dissolving theenteric coating polymer in a suitable solvent, preferably a solvent pairsuch as chloroform; methanol or isopropanol:ethyl acetate (1:1 w/w).Then, starting with the most concentrated solution, the tablets or beadsare sprayed in a coating pan under carefully controlled conditions, asdetermined by visual observation, until the desired film thickness,corresponding to the desired coating weight, is achieved. During thecoating operation, it has been observed that rapid aggregation of thetablets or the beads into multiplets. This, however, could be preventedby lightly dusting Talc U.S.P. over the overwetted tablets or beads.Many suitable enteric coating compositions meeting the requirements ofmy invention, as well as suitable alternative enteric coatingtechniques, are well known in the art. Thus, for example, a singlecoating solution, with a solids content as high as 10%, has been foundto be useful under appropriate conditions.

g. Dosage

The total amount of the composition required to be administered to anenzyme deficient patient will vary with the severity of the conditionand the amount of food ingested. Generally, for a pancreatic enzymedeficient patient from about 0.8 to 1.5 gms. of the final pancreaticenzyme composition are administered with each meal. This amounts toabout 2 about 3 capsules of small beads or tablets. In the presentlypreferred embodiment, each capsule comprises from about 0.425 to about0.475 gms. of digestive enzyme composition, whether in bead or tabletform.

h. Discussion of Tables

Table I below illustrates the deficiencies of currently available enzymecontaining products, which are ineffective or only partially effectiveto replace the missing enzymes in most pancreatic enzyme deficientpatients. As can be seen from the comparative data in that table, mostproducts on the market contain low levels of enzymes, they areespecially deficient in lipase, and their enteric coatings failadequately to protect the enzymes against gastric inactivation. Bycontrast, the compositions of the present invention are shown in Table Ibelow to comprise known high potency enzymes in an effective bindersystem. Moreover, they have properly balanced enteric coatings wherebyto avoid the inactivating effect of gastric acidity and pepsin.Therefore, the survival of the enzymes in active form is increasedsignificantly so that their bioavailability in the duodenum ismaximized.

Referring more particularly to Table I below, increased bioavailabilityis demonstrated for small tablets made of a composition of the presentinvention as described in Example 1. Thus, upon exposure to a simulatedgastric environment for two hours, the biological activity of thepreparations of this invention was well protected against gastricinactivation. Four of the nine commercially available enzymecompositions tested disintegrated within the first hour of exposure, andtheir enzymes were entirely inactivated. There were losses of at least40% in the activity of at least one enzyme in three of the other fivecommercial compositions. After exposure to simulated intestinal juices(slightly alkaline), the tablets of the present invention disintegratedwithin about 15 minutes and released the enzymes in their active state.In contrast, three of the five remaining commercial preparations, whichtested under identical conditions, failed to disintegrate in theintestinal environment within a reasonable time, and all haddisintegration times greater than those of the tablets of Example 1.Reasonable time is defined as the time that is required for the food tobe digested, transported and absorbed in the duodenum and upperintestine. The absorptive surfaces represent a segment of the upperintestine approximately 5 to 7 feet long. Transit of food through thissegment of the intestine by peristalsis under normal physiologicalconditions usually requires 45 to 60 minutes. Therefore, any entericcoated preparation that requires a longer disintegration time than 60minutes to release its enzyme content in this part of the upperintestine is considered to be defective, and hence to have littleusefulness to the patient.

Bioassays of the intestinal juices showed that between 74% and 84% ofthe initial activities of the enzymes of the tablets of Example 1 werepreserved during gastric exposure. By contrast, most of the commericalpreparations had lost at least 50% of the initial activity of at leastone of its enzymes. In fact, a comparison of the initial activitiesshown in Table I with the activities after 1 hour exposure to intestinalfluid (which followed the two-hour exposure to gastric fluid) shows thatonly the tablets of the present invention were able to maintain anddeliver a high concentration of active enzymes of all three enzymeswhose activities were tested, even though several of the commericalcompositions showed initial enzyme activities of at least one of theenzymes comparable to that of the tablets of Example 1.

Table II shows the performance of beads prepared in accordance withExample 9 after one hour exposure to gastric fluid followed by exposureto intestinal fluid. Note that complete disintegration, with delivery ofat least 95% of initial enzyme activity of each of the enzymes, tookplace within 20 minutes. Moreover, even after only 10 minutes exposureof the intestinal fluid, adequate delivery of active enzyme had alreadyoccurred (75.1% Protease; 85.5% Amylase; 100% Lipase).

Table III is self-explanatory and shows that isopropanol is a safe,fully compatible solvent for use with the enzymes in accordance with thepresent invention, whereas methanol has such poor compatibility that itis unsuitable. Acetone, chloroform and dioxane, while having somedeleterious effect on enzyme activity, are also suitable, although lessdesirable than isopropanol.

Further experimentation with freshly aspirated human gastric duodenaljuices revealed that the exogenously administered enzyme system of thepresent invention (tablets of Example 1) is compatible with humanduodenal juices and stabilizes the patient's pancreatic enzymes againsttemperature inactivation (Tables IV and V). That is, the enteric coatedenzyme tablets of Example 1 were resistant to gastric inactivation, andthe enzymes released in the duodenum were biologically active and werecompatible with the patient's enzyme system (Table IV). In addition, theexogenously administered enzyme system stabilized the patient's enzymesystem against temperature inactivation (Table V). This unexpectedbenefit resulted in longer digestion and better utilization of foods;and separate studies showed that the polymers polyvinylpyrrolidone andcellulose acetate phthalate (used as binders) stabilized the enzymesagainst temperature denaturation and self-digestion. Most conventionalpreparations, however, have been found to be substantially inactivatedwhen subjected to identical environmental conditions.

Referring to Table VI, disc gel electrophoretic studies of duodenaljuices aspirated after the administration of the enteric coateddigestive aid confirmed that the enzymes were released from the tabletsand beads into the duodenum and that the released enzymes werebiologically active. Thus, it can be seen that when the digestive enzymecompositions of this invention are employed, the biological activitiesof the enzymes are preserved and protected from gastric inactivation.Moreover, they are released in the duodenum within 5 to 30 minutes intheir active state.

Lipase is dependent on co-lipase for enzyme activity. As shown in TablesVII and VIII, it has been discovered during the course of myinvestigation that the presence of co-lipase in the compositions of thisinvention activates and stabilizes the lipase of pancreatic patients(Table VIII). Our clinical studies have shown that many pancreaticpatients, to some extent, are deficient in co-lipase, and thatco-lipase, stabilizes lipase against bile salt (Na taurocholate)inactivation. (Compare Table VII with Table VIII.) In the absence ofco-lipase, fat digestion is impaired. Therefore, an additional benefitof the preferred compositions of this invention is the availability anddelivery of an essential cofactor (co-lipase) that is necessary forlipid digestion. The manufacturing procedure outlined above preservesthe biological activity of co-lipase and the uniform enteric coatingensures that the co-lipase is delivered into the duodenum intact in itsbiologically active form.

j. Examples

The following specific examples are presented to further illustrate thecompositions and methods of this invention, without thereby limiting thescope thereof. Unless otherwise indicated, all percentages are byweight.

    ______________________________________                                        EXAMPLE 1                                                                      Phase 1                                                                              Pancreatic enzyme powder                                                      (5 × N.F. Pancreatin)                                                                           67.0%                                                 Polyvinylpyrrolidone    1.3%                                                  Starch (Sta-Rx 1500)    5.0%                                                  Sodium bicarbonate (anhydrous)                                                                        20.0%                                                 Citric acid             6.7%                                                                          100.0%                                                Solvent:                                                                      Isopropanol (anhydrous) 700 ml/kg blend                               Phase 2 (Enteric coating composition)                                                 Chloroform              66.4%                                                 Methanol (anhydrous)    15.4%                                                 Cellulose acetate phthalate                                                                           7.2%                                                  Talc #127 U.S.P.        7.3%                                                  FD & #5 yellow          1.0%                                                  Diethyl phthalate       2.7%                                                                          100.0%                                        ______________________________________                                    

The phase 1 composition was granulated, extruded and either tableted orprocessed into beads under the following conditions. The enzyme powderwas blended with polyvinylpyrrolidone, Starch 1500, sodium bicarbonateand citric acid in a Pony Mixer pan, with the slow addition of 700 ml/kgof isopropyl alcohol (anhydrous) for 9 to 15 minutes. The resultingblend was then segmented by means of a Stokes extruder, model EXKS-1,through openings of about 1 mm in diameter.

In preparing beads, the segmented particles were rounded off by means ofa marumerizer into spherical particles. The marumerizer speed was 430rpm. Time of marumerization was varied between 45 to 75 seconds. (Longermarumerization times are avoided as they were found to result in morecompact, dense particles. This in turn resulted in longer disintegrationtimes.) The resulting beads were about 10 - 12 mesh in size. Afterspheronization, the solvent was evaporated under controlled conditionsof low humidity, i.e., 5 to 10% relative humidity, and temperature,i.e., about 35° C.

To prepare small tablets, another portion of segmented particles wasdried in an oven at 35° C for approximately 40 to 48 hours. The driedgranules were sized through a 14 mesh screen. The segments which passedthrough the screen were then compressed on a Stokes BB 2 tablet machineemploying 3/16 deep cut tooling (or punches) to produce tablets about3/16 (4.8 mm) in diameter and about 0.155 (4 mm) thickness. Machinespeed was approximately 17 rpm.

The dried spherical granules or tablets were then coated with a pHsensitive enteric coating composition having the composition set forthabove (Phase 2) in an appropriate coating pan, e.g., one available fromthe Manesty Co., employing approximately 0.45 liters of coating solutionper kilogram of tablets and approximately 0.8 liters of coating solutionper kilogram of beads, until the appropriate film thickness wasachieved.

Additional examples of suitable Phase 1 compositions are set forth belowin Examples 2 - 9, while Example 9 also contains an illustration of asuitable alternative Phase 2 enteric coating composition, particularlysuitable for coating beads uniformly.

The Phase 1 compositions illustrated in Examples 2 through 8 are blendedin a mixing apparatus with the indicated solvent and then extruded intosegments by an extruder as described in Example 1, followed byprocessing into tablets or beads and enteric coating, also as describedin Example 1.

    ______________________________________                                        EXAMPLE 2                                                                      Phase 1                                                                             Pancreatic enzymes (5 × N.F. Pancreatin)                                                         90.0%                                                Polyvinylpyrrolidone K30 3.0%                                                 Avicel (crystalline cellulose having a                                        molecular weight in the range                                                 of 30,000 to 50,000)     7.0%                                                                          100.0%                                               Solvent:                                                                      Isopropanol 675 mg/kg blend                                            ______________________________________                                    

    ______________________________________                                        EXAMPLE 3                                                                      Phase 1                                                                             Pancreatic enzymes (5 × N.F. Pancreatin)                                                         89.0%                                                Polyvinylpyrrolidone K30 4.5%                                                 Sta-Rx (corn starch)     6.5%                                                                          100.0%                                               Solvent:                                                                      Isopropanol 625 ml/kg blend                                            ______________________________________                                    

    ______________________________________                                        EXAMPLE 4                                                                      Phase 1                                                                             Pancreatic enzymes (5 × N.F. Pancreatin)                                                         89.0%                                                Cellulose acetate phthalate                                                                            4.5%                                                 Sta-Rx (corn starch)     6.5%                                                                          100.0%                                        Solvent:                                                                      Acetone - Isopropanol (1:3) 700 ml/kg blend                                   ______________________________________                                    

    ______________________________________                                        EXAMPLE 5                                                                      Phase 1                                                                             Pancreatic enzymes (5 × N.F. Pancreatin)                                                         90.0%                                                Polyvinylpyrrolidone K30 2.0%                                                 Methocel 65 HG           1.0%                                                 Sodium carbonate (anhydrous)                                                                           4.0%                                                 Citric Acid (anhydrous)  3.0%                                                                          100.0%                                               Solvent:                                                                      Isopropanol 650-675 ml/kg blend                                        ______________________________________                                    

    ______________________________________                                        EXAMPLE 6                                                                      Phase 1                                                                             Pancreatic enzymes (5 × N.F. Pancreatin)                                                         90.0%                                                methocel 65 HG           1.0%                                                 Sodium carbonate (anhydrous)                                                                           4.0%                                                 Citric acid (anhydrous)  3.0%                                                 Polyvinylpyrrolidone K-30                                                                              2.0%                                                                          100.0%                                               Solvent:                                                                      Isopropanol 650-700 ml/kg blend                                        ______________________________________                                    

The citric acid (30 gm) and the polyvinylpyrrolidone (20 gm) areseparately dissolved in isopropanol (500 ml) before the addition to theabove blend.

    ______________________________________                                        EXAMPLE 7                                                                      Phase 1 Lactase derived from yeast                                                                           83.0%                                                  Methocel 65 HG         2.0%                                                   Sta-Rx (modified corn starch)                                                                        15.0%                                                                         100.0%                                                 Solvent:                                                                      Isopropanol 675-725 ml/kg blend                                      ______________________________________                                    

    ______________________________________                                        EXAMPLE 8                                                                      Phase 1 Enzyme blend*          87.02%                                                 Methocel 65 HG         2.0%                                                   Polyvinylpyrrolidone K-30                                                                            4.0%                                                   Sodium carbonate or bicarbonate                                                                      4.0%                                                   Citric acid (anhydrous)                                                                              3.0%                                                                          100.0%                                                 Solvent:                                                                      Isopropanol 650-750 ml/kg blend                                      ______________________________________                                         *Enzyme blend composition: Papain, Ficin, Bromelain (enzymes from plant);     Lipase, Cellulase, Protease, Amylase and Lactase (enzymes from                microorganisms).                                                         

    ______________________________________                                        EXAMPLE 9 Beads by Nonpareil Process                                           Phase 1                                                                             Pancreatic enzymes (5 × N.F. Pancreatin)                                                         80.0%                                                Polyvinylpyrrolidone K-30                                                                              6.5%                                                 Primojel (modified potato starch)                                                                      3.5%                                                 Nonpareil seeds (sugar seeds 20-30 mesh                                       size)                    10.0%                                                                         100.0%                                               Solvent:                                                                      Isopropanol 675-725 ml/kg blend                                        ______________________________________                                    

The composition of Example 9 (Phase 1) was processed in accordance withthe nonpareil process described above. The enteric coating was performedas described above using the Phase 2 enteric coating composition setforth below, to give a uniform coating weighing about 5.5% by weight ofthe final bead.

    ______________________________________                                         Phase 2 (Enteric coating composition)                                                 Cellulose acetate phthalate                                                                          8.0%                                                   Diethyl phthalate      2.0%                                                   Ethyl acetate          45.0%                                                  Isopropanol            45.0%                                                                         100.0%                                        ______________________________________                                    

The resulting beads were screened, giving a yield of about 90% by weightof beads in the 10-12 mesh size range.

                                      TABLE I                                     __________________________________________________________________________    COMPARATIVE EVALUATION OF COMMERCIAL DIGESTIVE                                ENZYME COMPOSITIONS WITH TABLET PRODUCT OF PRESENT INVENTION                                                                           DISINTE-                                                                      GRATION                                                                       TIME                                   PRODUCT                                                                              PROTEASE (μ/gm)                                                                         AMYLASE (μ/gm)                                                                       LIPASE (μ/gm)                                                                       (Minutes)            __________________________________________________________________________    INITIAL ACTIVITY                                                                              Panteric granules                                                                      186,200    29,600    7,970                           (μ/gm)       Pancrex V                                                                              87,000     7,000     260                                             Cotazyme 130,000    23,000    9,600                                           Entozyme 19,700     1,800     55                                              Donnazyme                                                                              14,700     2,160     915                                             Phazyme  13,600     2,280     630                                             Panteric 50,000     9,500     935                                             Viokase  238,000    27,000    10,800                                          Dactilase                                                                              37,650     6,500     880                                             Product of Example 1 *                                                                 211,000    29,600    11,400                          ACTIVITY AFTER  Pantoric Granules                                                                      NIL        NIL       NIL         25                  EXPOSURE TO GASTRIC                                                                           Pancrex V                                                                              72,000     6,500     250         N **                FLUID,pH 1.8, FOR                                                                             Cotazyme NIL        NIL       NIL         R ***               2 Hrs.          Entozyme 15,075     2,640     680         N                                   Donnazyme                                                                              8,850      1,440     1,030       N                                   Phazyme  4,100      440       370         N                                   Panteric 40,000     7,000     410         N                                   Viokase  NIL        NIL       NIL         5                                   Dactilase                                                                              NIL        NIL       NIL         60                                  Product of Example 1                                                                   177,000    24,900    8,425       N                   ACTIVITY AFTER  Panteric Granules                                                                      NIL        NIL       NIL         -- ****             EXPOSURE TO INTESTINAL                                                                        Pancrex V                                                                              62,000     5,000     130         60                  FLUID, pH 7.5, FOR                                                                            Cotazyme NIL        NIL       NIL         --                  1 Hr.           Entozyme 13,550     1,600     390         20                                  Donnazyme                                                                              5,780      2,030     400         25                                  Phazyme  8,375      725       110         40                                  Panteric 40,000     6,200     370         150                                 Viokase  NIL        NIL       NIL         --                                  Dactilase                                                                              NIL        NIL       NIL         --                                  Product of Example 1                                                                   151,000    21,200    7,100       15                  __________________________________________________________________________     * Example I, Phase I and II combined in tablet form. Tablet size: 3/16"       diameter and 0.155" thickness.                                                ** N = No disintegration                                                      *** R = Rapid disintegration                                                  **** -- = Not determined since disintegration had already occurred in         gastric fluid.                                                           

                  TABLE II                                                        ______________________________________                                        RATE OF ENZYME RELEASE                                                        FROM BEADS* INTO DUODENAL                                                     JUICE AFTER EXPOSURE TO GASTRIC JUICE                                                       PRO-                                                                          TEASE  AMYLASE    LIPASE                                        ______________________________________                                        1.  Initial Activity (μ/gm)                                                                    123,060  41,112   11,773                                                      (100%)   (100%)   (100%)                                  2.  Exposure to simulated                                                         gastric fluid for one                                                         hr. followed by exposure                                                      to simulated intestinal                                                       fluid, pH 7.5 at 37° C.                                                 5 min.         25,963   8,836    2,527                                                       (21.1%)  (21.5%)  (21.5%)                                     10 min.         92,445   35,135   13,196                                                      (75.1%)  (85.5%)  (100%)                                      15 min.         116,717  38,137   13,118                                                      (94.8%)  (92.8%)  (100%)                                      20 min.         118,023  39,411   13,040                                                      (96.0%)  (95.9%)  (100%)                                  ______________________________________                                         *Example 9, Phases I and II, in bead form.                                    Bead size: 10-12 mesh.                                                        Enteric coating = 5.5% w/w.                                              

                  TABLE III                                                       ______________________________________                                        COMPATIBILITY OF THE 5 × N.F.                                           PANCREATIN ENZYMES WITH SOLVENTS                                                            PRO-                                                                          TEASE  AMYLASE    LIPASE                                        ______________________________________                                        Untreated Control                                                                             100%     100%       100%                                      Methanol, Reagent Grade                                                                        16%      10%        45%                                      Isopropanol, N.F.                                                                             100%     100%       100%                                      Acetone, Reagent Grade                                                                         83%      75%        94%                                      Chloroform, Reagent Grade                                                                      96%      81%       100%                                      Dioxane, Reagent Grade                                                                         94%      76%       100%                                      ______________________________________                                         *Remaining enzymatic activities after granulation for 4 hours with the        respective solvents at 25° C.                                     

                                      TABLE IV                                    __________________________________________________________________________    COMATIBILITY OF THE TABLETS OF EXAMPLE I                                      WITH HUMAN DUODENAL FLUIDS AT 10° C.                                                    LIPASE                                                                             TRYPSIN                                                                             CHYMOTRYPSIN                                                                            AMYLASE                                                                              PROTEASE                                                                             RN-ase                    __________________________________________________________________________      Human Duodenal Fluid                                                                         91.8 212.7 178.5     5.9    23.8    16.9                       Digestive Enzyme Tablet of                                                                    204 170   170       23.4   33.7   308                         Example I                                                                     Digestive Enzyme Tablet of                                                    Example I Exposed to Human                                                                    323 368.6 286       39.3   58.4   326.3                       Duodenal Fluid for one Hr.                                                  __________________________________________________________________________     NOTE: Comparison of column 3 with the sum of columns 1 and 2 clearly show     that the digestive enzymes in the tablets of Example 1 are compatible wit     the human pancreatic enzymes (duodenal juice) and the biological              activities are complimentary to each other.                              

                                      TABLE V                                     __________________________________________________________________________    TEMPERATURE STABILITY OF THE TABLETS OF EXAMPLE I                             IN HUMAN GASTRIC AND INTESTINAL FLUIDS                                                             LIPASE                                                                             TRYPSIN                                                                             CHYMOTRYPSIN                                                                            AMYLASE                                                                              PROTEASE                     __________________________________________________________________________      Hormone* Stimulated                                                           Duodenal Fluid     383  680   595       7.33   6.42                           Hormone Stimulated                                                            Duodenal Fluid incubated                                                      at 37° C. for 40 min.                                                                     5.1  57.1  59.5      7.33   9.53                           Digestive Enzyme Tablet of Example I                                          in Phosphate Buffer, pH 7.4                                                                      204  170   170       23.4   33.7                           Digestive Enzyme Tablet of Example I                                          in Phosphate Buffer, pH 7.4                                                   incubated at 37° C. for 40 min.                                                           141  117   117       17.1   29.0                           Digestive Enzyme Tablet of Example I                                          Exposed to Human Gastric Fluid                                                (1 Hr.) followed by Duodenal Fluid                                            at 37° C. for 40 min.                                                                     478  382   306       45.4   62.4                           Sum of activities in Columns 2 and 4                                                             146.1                                                                              174.1 176.5     24.43  38.53                        __________________________________________________________________________     NOTE: Inspection of columns 1 vs. 2 and 3 vs. 4 show that exposure of the     respective enzyme solutions to elevated temperatures is detrimental to th     biological activities of the enzymes. If one sums up the activities of        column 2 with 4, column 6 is obtained. Comparison of columns 5 vs. 6          clearly show that the composition of the Tablets of Example I stabilizes      the biological activities of the duodenal juice against temperature           inactivation.                                                                 *Secretin and Cholecystokinin - Available for investigational use in          prepared ampules together with printed instructions for use from:GIH          Research Unit, Chemistry Department, Karolinska Institutet, S-104 01          Stockholm 60, Sweden.                                                    

                                      TABLE VI                                    __________________________________________________________________________    DISTINTEGRATION OF THE EXOGENOUSLY ADMINISTERED TABLETS                       OF EXAMPLE I IN THE HUMAN DUODENUM                                                                LIPASE                                                                             TRYPSIN                                                                             CHYMOTRYPSIN                                                                             AMYLASE                                                                              PROTEASE                                                                              PROTEIN              __________________________________________________________________________      Duodenal Aspirate 30 min.                                                     after tube insertion                                                                            0    0     0          10     0       144                    30 min. after administration                                                  of Digestive Enzyme Tablets of                                                Example I         0    0     0           6     0        25                    60 min. after Digestive                                                       Enzyme Tablets of Example I                                                                     1,521                                                                              4,973 4,973      16,120 2,178   426                  __________________________________________________________________________     NOTE: The appearance of biological activity after 60 min. (3) is proof        that the enzymes are released from the Tablets of Example I in the            duodenum, within 5-30 minutes after leaving the stomach.                 

                  TABLE VII                                                       ______________________________________                                        CO-LIPASE DEFICIENCY                                                          IN HUMAN DUODENAL JUICE                                                                                 WITHOUT                                             SUBJECT WITH TAUROCHOLATE*                                                                              TAUROCHOLATE                                        ______________________________________                                        R.P.    382               17,959                                              H.M.    120                 160                                               E.R.    5,663             43,250                                              E.R.     51               17,803                                              J.P.    2,446             14.693                                              J.C.     0                 3,289                                              ______________________________________                                         *Conc. of taurocholate 0.2 mg/ml                                              NOTE: Na-taurocholate inactivates lipase in the absence of co-lipase.    

                  TABLE VIII                                                      ______________________________________                                        ACTIVATION OF CO-LIPASE                                                       DEFICIENT HUMAN LIPASE                                                        BY THE ADDITION OF THE                                                        DIGESTIVE ENZYME                                                              TABLETS OF EXAMPLE I                                                                           WITH    WITHOUT                                                               TAUROC- TAUROC-                                                               HOLATE  HOLATE                                               ______________________________________                                        1.  Human Duodenal Aspirate                                                                           92       271                                          2.  Digestive Enzyme Tablets                                                      of Example I       204       102                                          3.  Digestive Enzyme Tablets                                                      of Example I Dissolved in                                                     Human Duodenal Aspirate                                                                          323       336                                          ______________________________________                                         NOTE: The increased activity in the presence of taurocholate (3) shows        that the free co-lipase in the Digestive Enzyme Tablets of Example I          activates and stabilizes the co-lipase deficient human lipase.           

What is claimed:
 1. In a process for preparing a digestive enzymecomposition for treating enzyme deficient mammals, said compositioncomprising (a) a concentrate of an enzyme from the group consisting ofthe pancreatic proteases, lipases, nucleases and amylase, theplant-derived digestive enzymes and the digestive enzymes derived frommicrobial sources in (b) a binder system comprising (i) at least about0.5 wt. % (based on the weight of the binder system plus enzymes) of abinder selected from the group consisting of polyvinylpyrrolidone,microcrystalline cellulose, cellulose acetate phthalate, methylcelluloseand alginic acid, and (ii) from zero to about 10 wt. % (based on theweight of the binder system plus enzymes) of a stabilizer selected fromthe group consisting of calcium carbonate, polyvinylpyrrolidone,cellulose acetate phthalate, methylcellulose, starch and modifiedstarches and alginic acid; and (c) from about 0.1% to about 30 wt. %,based on the weight of the total composite (enzyme plus binder systemplus disintegrant), of a disintegrant selected from the group consistingof citric acid, sodium carbonate, sodium bicarbonate, calcium carbonate,starch and modified starches, microcrystalline cellulose, and alginicacid; said process including the steps of blending said enzyme with saidbinder system and said disintegrant to form a composite and thereaftercoating said enzyme/binder system/disintegrant composite with anon-porous, pharmaceutically acceptable enteric coating which isinsoluble in the pH range of from about 1.5 to about 5 but is soluble inthe pH range of from about 6 to about 9, the improvement consisting ofcarrying out said process while avoiding the presence of water and saidblending step being performed in the presence of a single liquid phasecomprising an inert organic enzyme-compatible solvent.
 2. The process ofclaim 1 wherein said enzyme is selected from the group consisting ofTrypsin, Chymotrypsin, Chymotrypsin B, Pancreatopeptidase,Carboxypeptidase A, Carboxypeptidase B, Glycerol ester hydrolase,Phospholipase A₂, Sterol ester hydrolase, Ribonuclease,Deoxyribonuclease, α-Amylase, Papain, Chymopapain, Bromelain, Ficin,β-Amylase, Cellulase, β-Galactosidase, Subtilopeptidase A, andAspergillopeptidase A.
 3. The process of claim 1 wherein said enzymeconcentrate comprises an enzyme mixture, each milligram of whichcontains at least about 75 N.F. units of protease activity, at leastabout 75 N.F. units of amylase activity, at least about 10 N.F. units oflipase activity, and an effective amount of co-lipase.
 4. The process ofclaim 3 wherein said enzyme mixture further contains at least about 5International Units of ribonuclease activity per milligram.
 5. Theprocess of claim 1 wherein said stabilizer is present in an amount offrom about 0.1% to about 10% by weight.
 6. The process of claim 1wherein said binder is present in an amount of from about 1% to about 5%by weight.
 7. The process of claim 1 wherein said disintegrant ispresent in an amount of from about 2% to about 15% by weight.
 8. Theprocess of claim 1 wherein said binder is methyl cellulose.
 9. Theprocess of claim 1 wherein said binder is polyvinylpyrrolidone, whichalso acts as said stabilizer.
 10. The process of claim 1 wherein saidenteric coating is present in an amount of from about 2.5% to about 10%by weight of the entire composition.
 11. The process of claim 1 whereinsaid enteric coating comprises cellulose acetate phthalate and diethylphthalate.
 12. The process of claim 1 wherein said binder and saiddisintegrant are first dissolved in said solvent and the resultingsolution is then slowly added to said enzyme mixture.
 13. The process ofclaim 1 wherein said solvent is isopropanol.
 14. The process of claim 1,which includes the steps of granulating the resulting blend, extrudingit into segments, screening the segments and subsequently tableting theresulting granules at a sufficiently low compression pressure that thetemperature during compression is maintained in the range of from about15° to about 30° C.
 15. The process of claim 1 wherein said solvent isemployed in an amount of from about 600 to about 700 ml. per kg. ofblend of enzyme mixture plus binder plus disintegrant.
 16. The processof claim 1 wherein said blend is granulated, extruded through a screenwith uniform openings having a diameter in the range of between about0.5 mm and about 2 mm, and thereafter formed into spheres having a sizein the range of between about 8 and about 14 mesh by processing saidextruded pellets on a Marumerizer for from about 30 seconds to about 75seconds.
 17. The process of claim 16 wherein said Marumerization isperformed at a temperature not greater than about 20° C.
 18. The processof claim 1 wherein said blend is prepared by combining said disintegrantand said enzyme mixture in a suitable apparatus to form a uniform dryblend; said binder is separately dissolved in said solvent, and thecomposition is formed into spheres having a diameter in the range offrom about 8 to about 14 mesh by dusting said dry blend over nonpareilseeds tumbling and flowing in a coating pan, said seeds having beenwetted with said binder solution, with periodic addition of said solventto maintain the particles in a wetted but free-flowing state, until theseeds have been built to uniform spherical particles having diameterspredominantly in said range of from about 8 to about 14 mesh.
 19. Theprocess of claim 18 wherein said dusting operation is performed untilsaid diameters of said spheres are predominantly in the range of fromabout 10 to about 12 mesh.
 20. The process of claim 1 which furthercomprises applying an enteric coating to said tablets or spheres byspraying solutions of progressively lower concentrations of an entericcoating composition onto said tablets or spheres in a coating pan untilthe desired coating thickness is achieved.
 21. The process of claim 20wherein said enteric coating solution comprises cellulose acetatephthalate and diethyl phthalate in at least one solvent selected fromthe group consisting of acetone, methyl ethyl ketone, diacetone alcohol,ethylene glycol monoacetate, ethanol, chloroform, methanol, isopropanol,ethyl acetate, methylene chloride and benzene.
 22. The process of claim21 wherein said solvent is a mixture of equal parts by volume ofchloroform and methanol, or of isopropanol and ethylacetate.
 23. Adigestive enzyme composition prepared by the process of claim
 1. 24. Adigestive enzyme composition prepared by the process of claim
 14. 25. Adigestive enzyme composition prepared by the process of claim
 16. 26. Adigestive enzyme composition prepared by the process of claim
 18. 27. Amethod for treating digestive enzyme deficiency in mammals comprisingfeeding the mammal with each meal an effective amount of the compositionof claim
 25. 28. The method of claim 27, wherein from about 0.8 to about1.5 grams of said composition are fed with each meal.
 29. The method ofclaim 28 wherein said composition is in the form of spheres of a size inthe range of from about 8 to about 14 mesh.
 30. The method of claim 1wherein said inert organic enzyme-compatible solvent is a memberselected from the group consisting of isopropanol, methylene chloride,dioxane, tetrahydrofuran and acetone.